title: Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses.
abstract: We describe a STED microscope optimized for colocalization experiments with up to three colors. Two fluorescence labels are separated by their fluorescence lifetime whereas a third channel is discriminated by the wavelength of fluorescence emission. Since it does not require a second STED beam, separating by lifetime is insensitive to drift and thus optimally suited for colocalization analyses. Furthermore, we propose a setup having a second STED beam for long duration multicolor recording.
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Johanna
authors.initials: J
authors.lastname: Bückers
authors.forename: Dominik
authors.initials: D
authors.lastname: Wildanger
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Vicidomini
authors.forename: Lars
authors.initials: L
authors.lastname: Kastrup
authors.forename: Stefan W
authors.initials: SW
authors.lastname: Hell
mesh_terms: D000818:Animals; D002460:Cell Line; D003116:Color; D021621:Imaging, Three-Dimensional / Q000379:methods*; D034882:Lamins / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism
chemical_list: D034882:Lamins; D014404:Tubulin